CN105381092A - 一种治疗角膜炎的中药凝胶及其制备方法和用途 - Google Patents
一种治疗角膜炎的中药凝胶及其制备方法和用途 Download PDFInfo
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Abstract
本发明公开了一种治疗角膜炎的中药凝胶及其制备方法和用途，治疗角膜炎的中药凝胶的原料为金银花、密蒙花、防风、一叶萩、功劳木、蔓荆子、荆芥穗、丝瓜络、赤芍、鸡血藤、八角金盘。在制备时，加入了保湿剂、pH调节剂和缓冲剂，保湿剂为甘油或丙二醇，缓冲剂为枸橼酸钠/枸橼酸缓冲对、磷酸盐缓冲对、tris-盐酸缓冲对或甘氨酸-三乙胺缓冲对。本发明通过药物的透皮吸收，达到治疗角膜炎的目的，快速消除角膜炎引起的畏光、流泪、酸痛等症状，修补受损免疫系统，增强身体抵抗力，从根本上治疗角膜炎，直接作用于眼睛，没有毒副作用，为临床治疗角膜炎，缓解角膜炎引起的各种症状提供了一个新选择。
Description
技术领域
本发明涉及医药应用技术领域，具体是一种治疗角膜炎的中药凝胶及其制备方法和用途。
背景技术
单纯疱疹病毒性角膜炎是目前最严重的常见角膜病，近几年来有明显上升和加剧趋势。由于反复发作，重症病例增多，严重威胁视功能，故加强对本病的研究，已成为眼科亟待解决的课题。本病临床表现复杂，除了典型的树枝、地图及盘状角膜病灶形态外，还有一些不典型的临床改变，因此给诊断和治疗带来很大困难。各型不仅具有独特的临床特征，而其发病机理及治疗原则也不尽相同，染色各型之间是可以相互转化的，决定这种转化的因素非常复杂，除了与HSV的型和毒株及宿主的免疫功能(主要是细胞免疫)有关外，还与治疗方法有关(特别是皮质固醇的应用)。
角膜浅层有丰富的三叉神经末梢，故该病常有明显的刺激症状，有畏光、流泪、酸痛等。角膜本属透明，一旦有病，则其透明度发生改变，病人常主诉有视物模糊。该病一般沿三叉神经发病，病变部位侵犯较深，其感觉减退，但因炎症刺激角膜病变的邻近组织，因此刺激症状仍较明显。病毒性角膜炎，病程长，愈后且易复发。常可伴有葡萄膜反应，甚至出现虹膜睫状体炎、前房积脓，或继发青光眼，是临床上较为常见的致盲眼病之一。
目前临床上针对单纯疱疹病毒性角膜炎的治疗方法主要有：病灶清创术，适用于浅层型病例，其原理是通过物理或化学的方法来清除感染细胞和病毒，包括机械清创、化学清创、冷冻清创、光灭活疗法，病灶清创术只能清除感染细胞不能阻止病毒繁殖、可能损伤角膜上皮基底膜及基质层，促使病变向深部发展、冷冻术对HSV的活力无影响等问题；抗病毒药物，如碘苷、阿糖胞苷、三氟胸腺嘧啶核苷等，多用于单纯疱疹病毒性角膜炎后期治疗；皮质类固醇，抑制组织胺和毒性溶解酶的释放，减轻炎症反应及组织损害，减少疤痕形成，促进血管新生，但也存在损害机体免疫机制和损害角膜组织的风险；手术疗法，适用于深部溃疡、基质坏死性角膜炎合并穿孔者，单独依靠药物及保守治疗已难奏效的情况，包括结膜瓣遮盖术、前房穿刺术、板层或穿透角膜移植术，手术疗法存在临床手术风险。
近几年逐渐发展起来的新型治疗方法是应用免疫促进剂治疗，以及应用胶原酶抑制剂减少或防止溃疡的发生或发展。这些治疗方法与中药组方治疗方法存在一致性，均是通过调节或刺激机体免疫系统、提高体液或体细胞的免疫功能达到治疗目的。因为单纯疱疹病毒性角膜炎的发病机理是显性或陷性感染HSV从泪液中扩散至角膜、结膜、眼深部组织以及附属器，潜伏至神经末梢并激活，因此有效的治疗方法应通过提高细胞免疫能力、组织相容性抗原进行治疗。
传统中医理论认为，单纯疱疹病毒性角膜炎是由于实证热证、风热表实、脾虚湿热、肝胆火毒上攻所引发，应以清热解毒、疏散风热为纲，淡渗利湿、泄肝补肾为目，以求明目退翳之功效，从根本上调节内腑机能，调动机体和细胞免疫力，达到治愈单纯疱疹病毒性角膜炎的目的。
发明内容
本发明的目的在于提供一种治疗角膜炎的中药凝胶及其制备方法和用途。
为实现上述目的，本发明提供如下技术方案：
一种治疗角膜炎的中药凝胶，由以下按照重量份的原料组成：金银花30-40份、密蒙花30-40份、防风20-30份、一叶萩20-30份、功劳木10-20份、蔓荆子10-20份、荆芥穗8-12份、丝瓜络6-10份、赤芍6-10份、鸡血藤3-7份、八角金盘3-5份。
作为本发明进一步的方案：所述治疗角膜炎的中药凝胶由以下按照重量份的原料组成：金银花32-38份、密蒙花32-38份、防风22-28份、一叶萩22-28份、功劳木12-18份、蔓荆子12-18份、荆芥穗9-11份、丝瓜络7-9份、赤芍7-9份、鸡血藤4-6份、八角金盘3.5-4.5份。
作为本发明进一步的方案：所述治疗角膜炎的中药凝胶由以下按照重量份的原料组成：金银花35份、密蒙花35份、防风25份、一叶萩25份、功劳木15份、蔓荆子15份、荆芥穗10份、丝瓜络8份、赤芍8份、鸡血藤5份、八角金盘4份。
所述治疗角膜炎的中药凝胶的制备方法，具体步骤如下：
(1)按照重量份称取各原料；
(2)将原料药加水煎煮提取二次，第一次加水6-8倍量，煎煮2-4h，滤出药液，药渣继续加水6-8倍量，煎煮2-4h，滤出药液，药渣弃去；滤液合并浓缩至相对密度110-115得清膏，放冷，加入乙醇使其含醇量达70％，静置20-24h，滤取醇提上清液，回收乙醇得上清液并浓缩至相对密度为1.02-1.07的浓缩液，放冷，待用；
(3)取120-140份保湿剂加入至上述浓缩液中，搅拌均匀，静置至全部溶解，在搅拌下加入5-7份pH调节剂、6-8份缓冲剂，混合均匀，添加纯化水进行定容，搅拌均匀，静置24h得到中药凝胶成品。
作为本发明再进一步的方案：所述保湿剂为甘油或丙二醇。
作为本发明再进一步的方案：所述pH调节剂为三乙醇胺、氢氧化钠、碳酸氢钠、磷酸二氢钠、磷酸氢二钠、磷酸二氢钾、磷酸氢二钾、硼酸、硼砂、醋酸、醋酸钠、柠檬酸、柠檬酸钠、酒石酸、酒石酸钠、碳酸钠、碳酸钾、碳酸氢钠、碳酸氢钾、氢氧化钠、氢氧化钾、盐酸、硫酸、硝酸、磷酸中的一种或两种以上。
作为本发明再进一步的方案：所述缓冲剂为枸橼酸钠/枸橼酸缓冲对、磷酸盐缓冲对、tris-盐酸缓冲对或甘氨酸-三乙胺缓冲对。
作为本发明再进一步的方案：所述中药凝胶在制备治疗角膜炎药物中的应用。
与现有技术相比，本发明的有益效果是：
本发明通过药物的透皮吸收，达到治疗角膜炎的目的，快速消除角膜炎引起的畏光、流泪、酸痛等症状，修补受损免疫系统，增强身体抵抗力，从根本上治疗角膜炎，直接作用于眼睛，没有毒副作用，为临床治疗角膜炎，缓解角膜炎引起的各种症状提供了一个新选择。
具体实施方式
下面结合具体实施方式对本专利的技术方案作进一步详细地说明。
实施例1
一种治疗角膜炎的中药凝胶，由以下按照重量份的原料组成：金银花30份、密蒙花30份、防风20份、一叶萩20份、功劳木10份、蔓荆子10份、荆芥穗8份、丝瓜络6份、赤芍6份、鸡血藤3份、八角金盘3份。
所述治疗角膜炎的中药凝胶的制备方法，具体步骤如下：
(1)按照重量份称取各原料；
(2)将原料药加水煎煮提取二次，第一次加水6倍量，煎煮2h，滤出药液，药渣继续加水6倍量，煎煮2h，滤出药液，药渣弃去；滤液合并浓缩至相对密度110得清膏，放冷，加入乙醇使其含醇量达70％，静置20h，滤取醇提上清液，回收乙醇得上清液并浓缩至相对密度为1.02的浓缩液，放冷，待用；
(3)取120份甘油加入至上述浓缩液中，搅拌均匀，静置至全部溶解，在搅拌下加入5份三乙醇胺、6份枸橼酸钠/枸橼酸缓冲对，混合均匀，添加纯化水进行定容，搅拌均匀，静置24h得到中药凝胶成品。
使用方法：将本发明的中药凝胶滴于眼睑中，每次使用2-3ml，每日2次，早晚各一次。
实施例2
一种治疗角膜炎的中药凝胶由以下按照重量份的原料组成：金银花32份、密蒙花32份、防风22份、一叶萩22份、功劳木12份、蔓荆子12份、荆芥穗9份、丝瓜络7份、赤芍7份、鸡血藤4份、八角金盘3.5份。
所述治疗角膜炎的中药凝胶的制备方法，具体步骤如下：
(1)按照重量份称取各原料；
(2)将原料药加水煎煮提取二次，第一次加水6.5倍量，煎煮2.5h，滤出药液，药渣继续加水6.5倍量，煎煮2.5h，滤出药液，药渣弃去；滤液合并浓缩至相对密度113得清膏，放冷，加入乙醇使其含醇量达70％，静置21h，滤取醇提上清液，回收乙醇得上清液并浓缩至相对密度为1.03的浓缩液，放冷，待用；
(3)取125份丙二醇加入至上述浓缩液中，搅拌均匀，静置至全部溶解，在搅拌下加入5.5份氢氧化钠、6.5份磷酸盐缓冲对，混合均匀，添加纯化水进行定容，搅拌均匀，静置24h得到中药凝胶成品。
使用方法：将本发明的中药凝胶滴于眼睑中，每次使用2-3ml，每日2次，早晚各一次。
实施例3
一种治疗角膜炎的中药凝胶由以下按照重量份的原料组成：金银花35份、密蒙花35份、防风25份、一叶萩25份、功劳木15份、蔓荆子15份、荆芥穗10份、丝瓜络8份、赤芍8份、鸡血藤5份、八角金盘4份。
所述治疗角膜炎的中药凝胶的制备方法，具体步骤如下：
(1)按照重量份称取各原料；
(2)将原料药加水煎煮提取二次，第一次加水7倍量，煎煮3h，滤出药液，药渣继续加水7倍量，煎煮3h，滤出药液，药渣弃去；滤液合并浓缩至相对密度113得清膏，放冷，加入乙醇使其含醇量达70％，静置22h，滤取醇提上清液，回收乙醇得上清液并浓缩至相对密度为1.05的浓缩液，放冷，待用；
(3)取130份甘油或丙二醇加入至上述浓缩液中，搅拌均匀，静置至全部溶解，在搅拌下加入6份碳酸氢钠、6-8份tris-盐酸缓冲对，混合均匀，添加纯化水进行定容，搅拌均匀，静置24h得到中药凝胶成品。
使用方法：将本发明的中药凝胶滴于眼睑中，每次使用2-3ml，每日2次，早晚各一次。
实施例4
一种治疗角膜炎的中药凝胶由以下按照重量份的原料组成：金银花38份、密蒙花38份、防风28份、一叶萩28份、功劳木18份、蔓荆子18份、荆芥穗11份、丝瓜络9份、赤芍9份、鸡血藤6份、八角金盘4.5份。
所述治疗角膜炎的中药凝胶的制备方法，具体步骤如下：
(1)按照重量份称取各原料；
(2)将原料药加水煎煮提取二次，第一次加水7.5倍量，煎煮3.5h，滤出药液，药渣继续加水7.5倍量，煎煮3.5h，滤出药液，药渣弃去；滤液合并浓缩至相对密度114得清膏，放冷，加入乙醇使其含醇量达70％，静置23h，滤取醇提上清液，回收乙醇得上清液并浓缩至相对密度为1.06的浓缩液，放冷，待用；
(3)取135份甘油或丙二醇加入至上述浓缩液中，搅拌均匀，静置至全部溶解，在搅拌下加入6.5份磷酸二氢钠、7.5份甘氨酸-三乙胺缓冲对，混合均匀，添加纯化水进行定容，搅拌均匀，静置24h得到中药凝胶成品。
使用方法：将本发明的中药凝胶滴于眼睑中，每次使用2-3ml，每日2次，早晚各一次。
实施例5
一种治疗角膜炎的中药凝胶，由以下按照重量份的原料组成：金银花40份、密蒙花40份、防风30份、一叶萩30份、功劳木20份、蔓荆子20份、荆芥穗12份、丝瓜络10份、赤芍10份、鸡血藤7份、八角金盘5份。
所述治疗角膜炎的中药凝胶的制备方法，具体步骤如下：
(1)按照重量份称取各原料；
(2)将原料药加水煎煮提取二次，第一次加水8倍量，煎煮4h，滤出药液，药渣继续加水8倍量，煎煮4h，滤出药液，药渣弃去；滤液合并浓缩至相对密度115得清膏，放冷，加入乙醇使其含醇量达70％，静置24h，滤取醇提上清液，回收乙醇得上清液并浓缩至相对密度为1.07的浓缩液，放冷，待用；
(3)取140份甘油加入至上述浓缩液中，搅拌均匀，静置至全部溶解，在搅拌下加入7份酒石酸、8份枸橼酸钠/枸橼酸缓冲对，混合均匀，添加纯化水进行定容，搅拌均匀，静置24h得到中药凝胶成品。
使用方法：将本发明的中药凝胶滴于眼睑中，每次使用2-3ml，每日2次，早晚各一次。
对比例1
一种治疗角膜炎的中药凝胶由以下按照重量份的原料组成：金银花32份、密蒙花32份、防风22份、功劳木12份、蔓荆子12份、荆芥穗9份、丝瓜络7份、赤芍7份、鸡血藤4份、八角金盘3.5份。
所述治疗角膜炎的中药凝胶的制备方法，具体步骤如下：
(1)按照重量份称取各原料；
(2)将原料药加水煎煮提取二次，第一次加水6.5倍量，煎煮2.5h，滤出药液，药渣继续加水6.5倍量，煎煮2.5h，滤出药液，药渣弃去；滤液合并浓缩至相对密度113得清膏，放冷，加入乙醇使其含醇量达70％，静置21h，滤取醇提上清液，回收乙醇得上清液并浓缩至相对密度为1.03的浓缩液，放冷，待用；
(3)取125份丙二醇加入至上述浓缩液中，搅拌均匀，静置至全部溶解，在搅拌下加入5.5份氢氧化钠、6.5份磷酸盐缓冲对，混合均匀，添加纯化水进行定容，搅拌均匀，静置24h得到中药凝胶成品。
使用方法：将本发明的中药凝胶滴于眼睑中，每次使用2-3ml，每日2次，早晚各一次。
对比例2
一种治疗角膜炎的中药凝胶由以下按照重量份的原料组成：金银花38份、密蒙花38份、防风28份、一叶萩28份、功劳木18份、蔓荆子18份、荆芥穗11份、丝瓜络9份、赤芍9份、鸡血藤6份。
所述治疗角膜炎的中药凝胶的制备方法，具体步骤如下：
(1)按照重量份称取各原料；
(2)将原料药加水煎煮提取二次，第一次加水7.5倍量，煎煮3.5h，滤出药液，药渣继续加水7.5倍量，煎煮3.5h，滤出药液，药渣弃去；滤液合并浓缩至相对密度114得清膏，放冷，加入乙醇使其含醇量达70％，静置23h，滤取醇提上清液，回收乙醇得上清液并浓缩至相对密度为1.06的浓缩液，放冷，待用；
(3)取135份甘油或丙二醇加入至上述浓缩液中，搅拌均匀，静置至全部溶解，在搅拌下加入6.5份磷酸二氢钠、7.5份甘氨酸-三乙胺缓冲对，混合均匀，添加纯化水进行定容，搅拌均匀，静置24h得到中药凝胶成品。
使用方法：将本发明的中药凝胶滴于眼睑中，每次使用2-3ml，每日2次，早晚各一次。
实施例6
临床试验
(1)试验对象：采用本发明共治疗220例患者，其中男120例，女100例；年龄最大者71岁，年龄最小者15岁；
(2)诊断标准：
带状疱疹病毒感染，它是沿三叉神经末梢颞支分布，在眼睑的上睑及颞部皮肤程带状串珠状的疱疹，且以患眼的同侧为特点，一般皮肤病变决不超越中线。单纯疱疹病毒引起该病，常在感冒或高热中，一般有口角或鼻翼部出现单纯疱疹。牛痘病毒性角膜炎，有接种或接触过牛痘疫苗病史。以上三种在角膜常呈树枝状角膜炎，浸润后常发生溃疡，严重病例则常引起虹膜反应，虹膜睫状体炎，角膜后在裂隙灯检查下可见有灰白色的沉淀物，甚至有前房积脓。溃疡面扩大，可呈盘状或地图状，少数病例还可引起角膜穿孔，或角膜葡萄肿，继发性青光眼等，最终导致失明。
(3)治疗方法：将男120例，女100例病人平均分为五组，每组男24例、女20例，一组、二组和三组均为试验组，一组、二组和三组分别涂抹实施例1-实施例4制备得到的本发明的治疗角膜炎的中药凝胶，四组和五组为对照组，四组和五组分别涂抹对比例1和对比例2制备得到的中药凝胶，使用方法是将本发明的中药凝胶滴于眼睑中，每次使用2-3ml，每日2次，早晚各一次，十五天为一疗程。连续涂抹2个疗程后，统计五组的治疗结果，同时观察治疗期间患者出现的不良反应。
(4)治疗标准：
治愈：眼部刺激症状消失，充血消失，角膜浸润灶愈合，荧光素角膜染色不着色；
有效：眼部刺激症状明显减轻，角膜浸润灶范围缩小或浸润减轻，荧光素染色着色；
无效：眼部刺激症状明显，角膜浸润灶无缩小或扩大、加深，荧光素角膜染色明显着色。
(5)治疗结果：患者220例病例中涂药后多在十五天后见效，2-3个疗程治愈；其中治愈127例，有效61例，无效32例，总有效率达85.5％，治疗结果如表1所示：
表1：临床试验治疗结果
由表1可知，采用本发明的治疗角膜炎的中药凝胶进行治疗的平均有效率达到96.9％，采用对比例制备得到的中药凝胶的有效率只有68.2％，因此，本发明的药物之间具有协同增效作用，增加了中药凝胶的有效率。
典型病例：
(1)屈某某，男，57岁，右眼痛、流泪、怕光，当地医院诊断为病毒性角膜炎，曾给予无环鸟苷眼药水、泰利必妥眼药水治疗两周，病人说效果不明显。檢查：右眼混合充血，角膜见大片宽狹不等的浅表溃瘍，状如地图状，角膜部分轻度水肿，荧光素着色，角膜知觉纯，前房、虹膜未见明显异常。診断：右病毒性(HSV-1)角膜炎，给予本发明中药凝胶进行治疗，十天后，症状消失，溃疡愈合，留有轻度薄翳治愈。
(2)苏某，女，25岁，右眼HSV-1角膜炎复发三次，复发后用ACV(滴眼)治疗无效。检查：右混合充血，角膜中央部基质呈盘状水肿、混浊，并有皱褶，直径约5mm，角膜上皮粗糙无光泽，角膜后面沉着物附着于基质水肿区内皮细胞，角膜知觉饨，瞳孔较对侧略小，但未粘连。诊断：病毒性(HSV-1)角膜基质炎，当即给予本发明中药凝胶进行治疗，二十天后，症状消失，溃疡愈合，留有薄翳治愈。随访三年未复发。
本发明在大量的临床应用中发现其具有显著的治疗角膜炎的作用，现将主要的药效实验详述如下：
一、毒性实验：
取豚鼠100只(由合肥市实验动物和动物实验中心提供)，雌雄各半，体重为220-240g，分成5组，每组20只，给药前24h用脱毛剂在大鼠脊柱旁进行脱毛(约4cm×5cm)，将实施例1制备的中药凝胶多次均匀涂抹在第一组的脱毛区，将实施例2制备的中药凝胶多次均匀涂抹在第二组的脱毛区，将实施例3制备的中药凝胶多次均匀涂抹在第三组的脱毛区，将实施例4制备的中药凝胶多次均匀涂抹在第四组的脱毛区，将实施例5制备的中药凝胶多次均匀涂抹在第五组的脱毛区，每只动物按照7.5mL/kg剂量涂抹给药(经过反复试验，7.5mL/kg为该浸膏的最大给药量)，涂敷24h后，用温水擦拭残留的受试物。
给药后，各组大鼠连续观察14d，每天观察1～2次，并于给药前及给药后7d，14d称体质量，主要观察给药后动物的全身反应，包括皮毛、眼睛、口腔黏膜、呼吸、自主活动、肌张力等，有无颤抖、震颤、抽搐、流涎、腹泻、少动、精神萎靡不振、昏迷等毒性反应症状。记录有无死亡，若有死亡应立即进行尸检，若肉眼检查有明显异常时，进一步进行病理组织学检查。
给药当日，大鼠清醒，连续观察14d，各组大鼠皮毛均有光泽，眼睛无分泌物出现，口腔黏膜无溃烂、充血和红肿等刺激症状，能自主活动，行为无改变，均无烦躁等异常精神状态及全身中毒症状，14d后处死动物，解剖，肉眼观察各主要脏器，皮肤及脏器均无明显病理改变。
将给药处皮肤和不给药处皮肤取样后立即放入福尔马林中固定，常规采样，蜡埋，切片，制片，脱蜡，HE染色，封片，于显微镜下检查并摄影。镜下可见，给药处与不给药处皮肤表皮层与真皮层组织结构完整，间质无充血、水肿及炎性细胞浸润。
二、皮肤刺激性实验：
取豚鼠50只，雌雄各半，随机分为完整皮肤组和破损皮肤组，每组10只。用基质(75％乙醇)作为阴性对照，同体左右侧自身对比法。试验前24h将豚鼠背部剃毛，去毛范围左右各约3cm×3cm。破损皮肤组将脱毛部位用注射针头划“井”字并以出现渗血为度。将实施例1制备的中药凝胶多次均匀涂抹在第一组的脱毛区，将实施例2制备的中药凝胶多次均匀涂抹在第二组的脱毛区，将实施例3制备的中药凝胶多次均匀涂抹在第三组的脱毛区，将实施例4制备的中药凝胶多次均匀涂抹在第四组的脱毛区，将实施例5制备的中药凝胶多次均匀涂抹在第五组的脱毛区，每只动物按照1.5g涂抹于左侧去毛皮肤上，另一侧涂抹同样体积的基质对照。按上法每天给药1次，共7d，每次涂药部位一致。
观察指标和评价标准：
每次涂药前，在自然光线下观察并记录涂药部位有无红斑、水肿、色素沉着、出血点、皮肤粗糙或皮肤菲薄情况及其发生时间及消退时间，并按表2对红斑及水肿进行评分。末次涂药后48h和72h肉眼观察并记录涂抹部位有无红斑和水肿等情况。每次涂药后和次涂药后均按表2计算每一观察时间点各组受试物及阴性对照物皮肤刺激反应积分的平均分值，并按表3进行刺激强度评价。
表2皮肤刺激反应评分标准
表3皮肤刺激强度评价标准
给药期间各组动物一般状态、行为、体征等未见异常。各组动物给药部位均未见红斑及水肿等异常症状。各组动物给药及观察期间刺激性分值和刺激性强度评价结果见表4和表5。
表4各组药物对豚鼠正常皮肤刺激性反应情况
表5各组药物对豚鼠破损皮肤刺激性反应情况
由表4和表5可以看出，本发明的中药凝胶对正常皮肤和破损皮肤均没有刺激性。
三、动物学实验：
1.仪器：
显微镜XSP-BM19A：上海光学仪器六厂生产；TGL-16G高速离心机：上海安宁科学仪器厂；-86℃海尔超低温冰箱；小鼠灌胃器；JEM21230透射电镜；Bio-Rcd2550酶标仪。
2.试剂与药品：
本发明实施例1制备的中药凝胶；单纯疱疹病毒I型SM44株；补体致敏酵母菌冻干试剂及未致敏酵母菌冻干试剂；水合氯醛溶液、0.25％戊二醛溶液、10％福尔马林溶液等。
3.实验动物：
鼠龄为3-5周，体质量18-22g的健康清洁级雌性小鼠60只。由合肥市实验动物和动物实验中心提供，动物房温度(20.0±3.0)℃，自然光照，喂饲以灭菌颗粒饲料，饮用消毒处理的自来水，动物自由摄食饮水，各组小鼠在实验期间按组分笼饲养。
4.造模方法：
小鼠适应环境1周后，同时予每只小鼠双眼点0.25％氯霉素眼药水，2次/d。第二周开始将实验小鼠用水合氯醛0.06m1腹腔注射麻醉后，置于显微镜下，用25号无菌手术刀片交叉划伤右眼角膜上皮层，将5μl含有1×107PFU病毒的无菌最小基础培养基点于右眼角膜表面并轻轻按摩鼠眼睑30s。造模前后分别予小鼠角膜荧光素钠染色并用裂隙灯显微镜检查角膜改变。小鼠角膜感染病毒后18h荧光素钠染色，裂隙灯显微镜下检查角膜改变。发现原发角膜感染表现为急性上皮型角膜炎，小鼠角膜出现点状、片状上皮缺损，荧光素钠染色阳性，单纯疱疹病毒性角膜炎小鼠造模成功。
5.分组及给药：
将造模成功的小鼠45只随机分为三组，分为模型组、治疗组和空白组，每组15只。造模后第2天开始给药，将本发明中药凝胶以及生理盐水等分别滴入各组动物眼结膜囊内，给药后使眼睛被动闭合3-5s。实验第1-3天，每1-2h用药1次，每次1滴，每天8-10次；以后每天6-8次，每次1滴，连续给药三周。
6.小鼠状况的观察与红细胞C3b受体活性表达的测定：
小鼠角膜接种HSV-I后用2％荧光素钠生理盐水溶液滴双眼染色，闭眼1min，裂隙灯显微观察角膜病变，用药后第7、14、21天，以2％荧光素钠生理盐水溶液滴双眼染色，闭眼1min，裂隙灯显微观察角膜病变，角膜上皮点状、树枝状、地图状着色及基质混浊。
病变程度判断标准：以0至4计分；
0分：无着色和混浊；
1分：着色和混浊的范围为角膜面积的1/4以内；
2分：着色和混浊的范围为角膜面积的1/4～1/2之间；
3分：着色和混浊的范围为角膜面积的1/2～3/4之间；
4分：着色和混浊的范围为角膜面积的3/4以上。
末次给药12h后，将小鼠水合氯醛腹腔麻醉，眼球放入10％甲醛溶液固定，常规石蜡包埋。摘眼球取血，血液放入抗凝试管，离心去上清后，低温冰箱保存，进行红细胞C3b受体花环率实验。
7.结果：
(1)各组小鼠角膜病变综合疗效比较，结果见表6。
表6各组小鼠病变综合疗效评分比较(x±s)
| 组别 | 造模前 | 7d | 14d | 21d |
| 空白组 | 0±0 | 0±0 | 0±0 | 0±0 |
| 模型组 | 0±0 | 2.45±0.99 | 2.71±1.15 | 2.91±0.53 |
| 治疗组 | 0±0 | 1.23±0.54 | 0.58±0.34 | 0.12±0.07 |
从表6可以看出，采用本发明的中药凝胶在角膜病变方面与模型组相比，角膜病变程度明显减轻，证明对角膜病变具有良好的改善。
(2)各组小鼠用药后红细胞C3b受体活性水平的比较，结果见表7。
表7各组小鼠用药后红细胞C3b受体活性水平比较(x±s)
| 组别 | 红细胞C3b受体花环率(％) |
| 空白组 | 20.12±1.47 |
| 模型组 | 16.35±1.08 |
| 治疗组 | 21.78±1.51 |
红细胞不仅能黏附和运转有害的免疫复合物，而且能浓缩抗原，将抗原传递给T淋巴细胞，增强和活化T淋巴细胞的免疫反应，从表7可以看出，经过造模后，小鼠红细胞C3b受体花环率显著降低，采用本发明的治疗组对模型治疗后能够显著提高红细胞C3b受体花环率。
本发明通过药物的透皮吸收，达到治疗角膜炎的目的，快速消除角膜炎引起的畏光、流泪、酸痛等症状，修补受损免疫系统，增强身体抵抗力，从根本上治疗角膜炎，直接作用于眼睛，没有毒副作用，为临床治疗角膜炎，缓解角膜炎引起的各种症状提供了一个新选择。
上面对本专利的较佳实施方式作了详细说明，但是本专利并不限于上述实施方式，在本领域的普通技术人员所具备的知识范围内，还可以在不脱离本专利宗旨的前提下作出各种变化。
Claims (8)
1.一种治疗角膜炎的中药凝胶，其特征在于，由以下按照重量份的原料组成：金银花30-40份、密蒙花30-40份、防风20-30份、一叶萩20-30份、功劳木10-20份、蔓荆子10-20份、荆芥穗8-12份、丝瓜络6-10份、赤芍6-10份、鸡血藤3-7份、八角金盘3-5份。
2.根据权利要求1所述的治疗角膜炎的中药凝胶，其特征在于，由以下按照重量份的原料组成：金银花32-38份、密蒙花32-38份、防风22-28份、一叶萩22-28份、功劳木12-18份、蔓荆子12-18份、荆芥穗9-11份、丝瓜络7-9份、赤芍7-9份、鸡血藤4-6份、八角金盘3.5-4.5份。
3.根据权利要求1所述的治疗角膜炎的中药凝胶，其特征在于，由以下按照重量份的原料组成：金银花35份、密蒙花35份、防风25份、一叶萩25份、功劳木15份、蔓荆子15份、荆芥穗10份、丝瓜络8份、赤芍8份、鸡血藤5份、八角金盘4份。
4.一种如权利要求1-2任一所述的治疗角膜炎的中药凝胶的制备方法，其特征在于，具体步骤如下：
（1）按照重量份称取各原料；
（2）将原料药加水煎煮提取二次，第一次加水6-8倍量，煎煮2-4h，滤出药液，药渣继续加水6-8倍量，煎煮2-4h，滤出药液，药渣弃去；滤液合并浓缩至相对密度110-115得清膏，放冷，加入乙醇使其含醇量达70%，静置20-24h，滤取醇提上清液，回收乙醇得上清液并浓缩至相对密度为1.02-1.07的浓缩液，放冷，待用；
（3）取120-140份保湿剂加入至上述浓缩液中，搅拌均匀，静置至全部溶解，在搅拌下加入5-7份pH调节剂、6-8份缓冲剂，混合均匀，添加纯化水进行定容，搅拌均匀，静置24h得到中药凝胶成品。
5.根据权利要求4所述的治疗角膜炎的中药凝胶的制备方法，其特征在于，所述保湿剂为甘油或丙二醇。
6.根据权利要求4所述的治疗角膜炎的中药凝胶的制备方法，其特征在于，所述pH调节剂为三乙醇胺、氢氧化钠、碳酸氢钠、磷酸二氢钠、磷酸氢二钠、磷酸二氢钾、磷酸氢二钾、硼酸、硼砂、醋酸、醋酸钠、柠檬酸、柠檬酸钠、酒石酸、酒石酸钠、碳酸钠、碳酸钾、碳酸氢钠、碳酸氢钾、氢氧化钠、氢氧化钾、盐酸、硫酸、硝酸、磷酸中的一种或两种以上。
7.根据权利要求4所述的治疗角膜炎的中药凝胶的制备方法，其特征在于，所述缓冲剂为枸橼酸钠/枸橼酸缓冲对、磷酸盐缓冲对、tris-盐酸缓冲对或甘氨酸-三乙胺缓冲对。
8.一种如权利要求1-3任一所述的中药凝胶在制备治疗角膜炎药物中的应用。
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